A study of organisms in soil samples from southern Indiana

Butler University Botanical Studies Volume Article A study of organisms in soil samples from southern Indiana which inhibit the growth of Escherichia coli and Staphylococcus aureus Doris Colligan Follow this and additional works at: https://digitalcommons.butler.edu/botanical The Butler University Botanical Studies journal was published by the Botany Department of Butler University, Indianapolis, Indiana, from 1929 to 1964 The scientific journal featured original papers primarily on plant ecology, taxonomy, and microbiology Recommended Citation Colligan, Doris (1949) "A study of organisms in soil samples from southern Indiana which inhibit the growth of Escherichia coli and Staphylococcus aureus," Butler University Botanical Studies: Vol , Article Retrieved from: https://digitalcommons.butler.edu/botanical/vol9/iss1/3 This Article is brought to you for free and open access by Digital Commons @ Butler University It has been accepted for inclusion in Butler University Botanical Studies by an authorized editor of Digital Commons @ Butler University For more information, please contact digitalscholarship@butler.edu Butler University Botanical Studies (1929-1964) Edited by Ray C Friesner The Butler University Botanical Studies journal was published by the Botany Department of Butler University, Indianapolis, Indiana, from 1929 to 1964 The scientific journal featured original papers primarily on plant ecology, taxonomy, and microbiology The papers contain valuable historical studies, especially floristic surveys that document Indiana’s vegetation in past decades Authors were Butler faculty, current and former master’s degree students and undergraduates, and other Indiana botanists The journal was started by Stanley Cain, noted conservation biologist, and edited through most of its years of production by Ray C Friesner, Butler’s first botanist and founder of the department in 1919 The journal was distributed to learned societies and libraries through exchange During the years of the journal’s publication, the Butler University Botany Department had an active program of research and student training 201 bachelor’s degrees and 75 master’s degrees in Botany were conferred during this period Thirty-five of these graduates went on to earn doctorates at other institutions The Botany Department attracted many notable faculty members and students Distinguished faculty, in addition to Cain and Friesner , included John E Potzger, a forest ecologist and palynologist, Willard Nelson Clute, co-founder of the American Fern Society, Marion T Hall, former director of the Morton Arboretum, C Mervin Palmer, Rex Webster, and John Pelton Some of the former undergraduate and master’s students who made active contributions to the fields of botany and ecology include Dwight W Billings, Fay Kenoyer Daily, William A Daily, Rexford Daudenmire, Francis Hueber, Frank McCormick, Scott McCoy, Robert Petty, Potzger, Helene Starcs, and Theodore Sperry Cain, Daubenmire, Potzger, and Billings served as Presidents of the Ecological Society of America Requests for use of materials, especially figures and tables for use in ecology text books, from the Butler University Botanical Studies continue to be granted For more information, visit www.butler.edu/herbarium Primary Red, PiDe Secondary 706 mm 146mm 575 135 609 61.1 195 247 249 222 232 520 428 415 323 273 f' ­ ~ A STUDY OF ORGANISMS IN SOIL SAMPLES FROM SOUTHERN INDIANA WHICH INHIBIT THE GROWTH OF ESCHERICHIA COLI AND STAPHYLOCOCCUS A UREUSl By -._ DORIS COLLIGAN The purpose of this investigation was to determine the numbers and kinds of organisms-bacteria, actinomycetes, or fungi-found in certain soils which inhibit the growth of two test organisms Soil samples collected from beneath different species of trees in wooded areas were used to ascertain what correlation there might be between the number and kind of inhibitors found and the kind of soil from which they were isolated This ecological aspect of the study of soil organisms which pro­ duce antibiotic substances has not been emphasized up to the present time by any of the workers studying antibiotics and the organisms which produce them, but it would seem to be of some value to have an idea where the largest number of most active inhibitors are found Certainly any study of soil organisms showing inhibition to other micro-organisms is valuable since notations like the following in a paper by Hoogerheide (5) are common in botanical literature: "Waksman and Woodruff isolated from the soil in 1940 a new chromogenic species of Actinomyces which showed strong antagonistic properties toward all bacteria belonging to both the Gram-positive and Gram-negative types This new species was later described as Actinomyces Gnfibioficlls." Much work remains to be done in the search for new antibiotic sub­ stances to combat plant and animal diseases PROCEDURE Soil samples were collected in October, 1947, from wooded areas in southern Indiana in regions of unglaciated clay soil: at Cornus Ridge, in Brown County, and at Stoney Lonesome, in Bartholomew County The samples of soil were collected under seven different t A portion of a thesis submitted in partial fulfillment" of the requirements for the graduation honor magna cum laude, Department of Botany, Butler University kinds of trees From the Comus Ridge area the trees used were Pinus strobus, Liriodendl'on tulipifera, and Quercus montana; while at Stoney Lonesome soil was collected under Ace'!' saccha.rum, Carya ovata, Liriodendron ttdipifera, Juglans nigra, and Q1/,erws alba Two samples were taken from beneath each tree, one from the surface soil and one at a depth of two or three inches, so that a com­ parison might be obtained between the number of inhibitors at the two depths In collecting the surface soil, care was taken to avoid getting only the humus material on top of the ground \i\lhen the subsurface sample was taken, the surface earth was turned back first and then the soil taken from two or three inches down An alcohol lamp was used to flame the trowel used in taking samples so that organisms would not be carried from one area to another, and the soil was put into paper bags, numbered according to location and lettered "A" for surface soil or "B" for subsurface soil Two methods of proceeding to fine! organisms in the soil which would inhibit the growth of the test organisms, Escherichia coli and Sta,phylococct/.s aureus, were tried Up to a certain point both methods were parallel: one gram of soil was allowed to stand overnight in 10 cc of sterile distilled water One cc of this was again diluted in 10 cc of sterile distilled water, making a dilution of 1-100 One cc of the 1-100 dilution of soil was plated with sterile pipettes into each of 14 sterile petri dishes Seven of these petri dishes contained agar at at suitable pH for the growth of bacteria and actinomycetes and the other seven were poured with agar suitable for the growth of fungi Following are the formulae for the media used: Medil1m I (for bacteria and actinomycetes) : 10 5 Dextrose Beef Extract NaCI Peptone Agar (2%) Water grams grams grams grams 20 grams liter ~\'1edium II (for fungi, pH lowered to about 4.5 by the addition of corn steep liquor) : 30 grams grams 0.5 grams Dextrose _ :NaNo• _ K!HPO 10 ~gSO, ZnSO, '\gar (2%) Water Corn Steep Liquor These plates were incubated at growth of the fungi being somev and actinomycetes At this point the two metho' was that of inoculating tubes of pouring this on top of the soil sa cient growth on the plates Zo aureus would then be visible arOl produced a substance inhibitory proved unsatisfactory, however, ascertain whether a colony was it was extremely dif ficult to obt; doing the inhibiting due to the f